Universal rapid diagnostic test reader with trans-visual sensitivity

ABSTRACT

A universal rapid diagnostics test reader is disclosed and described herein that includes a set of control electronics, a digital camera component, an illumination component, a housing component, and a rapid diagnostics test tray, wherein the tray can hold at least one rapid diagnostics test having a shape and a size in a fixed position relative to the digital camera component and the illumination component, and wherein the reader can accommodate more than one different rapid diagnostics test. Methods are also disclosed that include: providing at least one first rapid diagnostics test having a first physical size, first feature and first format; providing at least one second rapid diagnostics test having a second physical size, second feature and second format; inserting the first rapid diagnostics test in a universal rapid diagnostics test reader; analyzing the first rapid diagnostics test using the universal rapid diagnostics test reader; removing the first rapid diagnostics test from the reader; inserting the second rapid diagnostics test in a universal rapid diagnostics test reader without any mechanical adjustments of the reader or without the use of any additional parts or additional inserts; and analyzing the second rapid diagnostics test using the universal rapid diagnostics test reader.

This United States Utility Application claims priority to U.S.Provisional Application Ser. No. 61/889,821 entitled “Universal RapidTest Reader for Lateral Flow Immunoassays with High Sensitivity”, U.S.Provisional Application Ser. No. 61/845,742 entitled “Flash-controlled,Wireless, Lensless, Universal Rapid Diagnostics Test (RDT) Reader”, U.S.Provisional Application Ser. No. 61/899,116 entitled “Non-uniform FlashIllumination Based focusing Method for the Imaging of Targets that areuniformly illuminated”, which are commonly-owned and incorporated hereinby reference in their entirety.

FIELD OF THE SUBJECT MATTER

The current application is related to a universal rapid diagnostic testreader with trans-visual sensitivity.

BACKGROUND

Rapid diagnostic tests (RDTs) play an important and growing role in thecontinuum of care worldwide. Administered either at the point of care indoctors offices, hospitals, urban and remote clinics, or by ambulatoryhealth workers and providing immediate results these tests contribute toimproved access, lower cost, and better quality healthcare. Anincreasing number of RDTs are available for home use by patients and thegeneral public for testing of acute and chronic conditions. The dominanttechnology used for RDTs is Lateral Flow Immuno-Chromatographic assay(LFI) and with the worldwide annual value of LFI tests and services of$18 B according to BCC Research. RDTs are also available in othervariations of immunoassays, such as fluorescent LFIs, flow-through, anddipstick tests. In fact, contemplated embodiments described here areapplicable to any RDT using a change of the optical properties as themechanism of action.

As valuable as RDTs are, they can be less reliable and accurate, becausethey are typically read visually, and therefore, are subject to humanerror [1-19]. These inherent errors can be substantially alleviatedthrough the use of electronic readers originally developed by ESE GmBHand today available from a number of sources [21, 22]. They aretypically desktop instruments for laboratory use, can be rather largeand heavy and can cost thousands of dollars. Recently, significantprogress in the state-of-the-art technology was achieved by ProfessorAydogan Ozcan and his research group at UCLA using a smartphone as thetechnology platform. In addition, they developed a reader [17, 2](hereafter Mudanyali reader) with the following advantages: a) small,handheld and light (˜2.3 oz), b) sensitive and accurate withtransmission or reflection readout mode, c) impervious to ambientlighting conditions, d) automated test readout with electronic datacapture and telemetry using smartphone communication capabilities, e)centralized data collection with geomapping capabilities and interfacesto health information systems, and f) low cost achieved by piggybackingon the enormous production volume of smartphones.

Despite the advantages, there are opportunities available for theseconventional readers to be improved. For instance, conventional systemsachieve low cost by using a smartphone which is inserted into a readerbody that provides RDT illumination, ambient isolation, and cassettehousing. However, different models of smartphones from a singlemanufacturer or even more from a variety of vendors all have differentmechanical dimensions, and they wouldn't fit into a body designed forone specific smartphone model. This precludes users from using their ownsmartphone for the reader: they have to buy another dedicated smartphonewhich is a significant cost increase.

Readers require sources of illumination and associated controlelectronics and battery housed outside the smartphone. In Mudanyali'sconventional reader, the control is provided by the software applicationin the smartphone via a cable which plugs into the smartphone micro USBpower connector. External cabling adds to the cost and reducesreliability; besides, many smartphones do not have the capability foroutbound control through their power connector. Also, Mudanyali's readerdescribes a power source disposed in the attachment such that theself-powered reader can be controlled via a physical button located onthe attachment. This operation fully depends on operator's ability touse the reader and increases the complexity of operation. It would beideal if contemplated readers and systems corrected many of the beforementioned deficiencies of the prior art.

Moreover, Mudanyali's reader is capable of accommodating different teststypes using special customized-trays per cassette type. Therefore, itdoesn't provide a universal solution to image any test withoutadditional mechanical components. A universal reader should be readilyable to work with a significant number of different test cassetteswithout a need for any mechanical adaptation or additional mechanicalcomponents.

Recently another implementation of a smartphone-based reader has beendisclosed [23], which depends on the optimized Rayleigh/Mie scatterdetection by taking into consideration the optical nitrocellulosemembrane and gold nanoparticles on rapid tests. For each test type, thisapproach requires a complicated and precise calibration procedure todetermine the optimum angles of illumination that minimize the Miescattering from the membrane while maximizing the Rayleigh scatterdetection from the gold nanoparticles on and inside the membrane. Due tothe significant variation between different RDT types and also thevariation within the samples of same RDT type in terms of use ofcomponents (e.g., membranes and nanoparticles) and position/orientationof membrane and cassettes, successful implementation of this concept ona portable unit is quite challenging and not feasible. For instance, thecoefficient of variation (CV) exceeds 50% in some of their measurementson quantitative tests [2]. This reader variation is generally notacceptable even in qualitative measurements. This alignment-dependentapproach may be useful for research purposes on the analysis ofcustom-made immunoassays using advanced optical imaging setups thatincludes a precise automated scanning stage and other opto-mechanicalcomponents.

Note that although the work here was focused on smartphone-based RDTreaders as the most advantageous architecture many of the technologiesdescribed herein apply equally well to any reader architecture based ondigital imaging.

SUMMARY OF THE SUBJECT MATTER

A universal rapid diagnostics test reader is disclosed and describedherein that includes a set of control electronics, a digital cameracomponent, an illumination component, a housing component, and a rapiddiagnostics test tray, wherein the tray can hold at least one rapiddiagnostics test having a shape and a size in a fixed position relativeto the digital camera component and the illumination component, andwherein the reader can accommodate more than one different rapiddiagnostics test.

Methods are also disclosed that include: providing at least one firstrapid diagnostics test having a first physical size, first feature andfirst format; providing at least one second rapid diagnostics testhaving a second physical size, second feature and second format;inserting the first rapid diagnostics test in a universal rapiddiagnostics test reader; analyzing the first rapid diagnostics testusing the universal rapid diagnostics test reader; removing the firstrapid diagnostics test from the reader; inserting the second rapiddiagnostics test in a universal rapid diagnostics test reader withoutany mechanical adjustments of the reader or without the use of anyadditional parts or additional inserts; and analyzing the second rapiddiagnostics test using the universal rapid diagnostics test reader.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows (a) Universal RDT reader attachment that can repeatedlyattached/detached to a cell-phone (b) PCB is enclosed within the readerattachment.

FIG. 2 shows (a-b) Different schematic views of proposed universal RDTreader prototype installed on an Android phone (Motorola Defy XT 535).This compact attachment can be repeatedly attached/detached to thesmartphone body without the need for fine alignment and anymodification. (c) Mechanical body of the reader attachment will alsoensure the isolation to the RDTs that are loaded to the smartphonereader attachment using a universal test tray. With its slip-free gripdesign, it allows the user to conveniently hold it even in the fieldsettings. It includes a single USB port with a photo-sensor tocommunicate with both smartphone and the PCB board. Having anunconventionally large field-of-view of ˜45 mm×85 mm, this attachmentwill also allow the user to acquire images of other objects of interest,such as user ID card (e.g., military) and RDT pouch with type/lotnumbers, while the universal RDT tray is retracted from the baseattachment assembly. (d) Reader attachment uses inexpensive opticalcomponents, i.e., a plano-convex lens (optional) and multiple diffusednarrow-band LEDs embedded on a single PCB that also carries circuitryfor the data and power communication with the smartphone andrechargeable flat or AA batteries, respectively. (e) Schematic diagramof the proposed optical geometry is shown. (f) Flexible lens assembly isdemonstrated. Utilizing an external diffuser and a narrow band-pass (<30nm) filter, flash light of the smartphone can be used to potentiallysupport or mimic the LED illumination. Without any modification on thearchitecture of the main mechanical body (c), lens holder can bemodified to hold the additional diffuser and band-pass filter as shownin (f).

FIG. 3 shows one of the major objectives of this development was toenable universal operation using the RDT reader, such that it canaccommodate a broad range of RDTs without a need for using multiplecustomized RDT trays and any modification on the reader attachment. Herewe propose a single universal RDT tray (b) that will accommodatemultiple (at least seven) RDT types. Starting with a selection of sevenRDTs we conceptually designed the universal tray that allows the user topress-fit the RDTs based on a variety of orientations and heights.Following the identifiers printed on this universal tray, user caneasily decide on the direction and orientation of the particular RDT tobe inserted and load the universal tray into the base reader attachmentas shown in FIG. 1(a-c). This design feature overcomes the need of usingindividual trays and attachment customized for each RDT, having animportant advantage especially in the field settings. It will alsoprovide significant reduction on material cost. In (a), different viewsof the universal tray loaded with different RDTs that will tested withinthe proposal timeline are shown. RDTs that will be examined using theproposed RDT reader in this proposal: (1) Oratect Plus (Branan MedicalCorp.) for drug screening, (2) ThyroChek (Thyrometrix, Inc.) forThyrotropin screening, (3) BioThreat Alert™ Test (Tetracore, Inc.) foranthrax diagnostics, (4) Cardiac Panel Test (MP Biomedicals, LLC) toscreen cardiac markers, (5) OptiMAL-IT Malaria RDT (Bio-Rad), (6)Uni-Gold™ HIV RDT (Trinity Biotech), (7) a custom hand held assay forbacterial antigen detection (developed by ECBC).

FIG. 4 shows (a, b, c) Photo of the Rapid Test Reader prototypesfabricated using a 3-D printer. (a) A prototype with a slip-on universalRDT tray can accommodate up to 7 different RDTs (b) A single-piecedesign that with relatively smaller dimensions can accommodate and fullyenclose up to 7 different RDTs (c) A third single-piece design that canfully enclose and accommodate up to 5 different RDTs has significantlysmaller dimensions and weight as well as smooth edges and corners. (d,e, f) Different types of RDTs, e.g., Uni-Gold™ HIV RDT (d), CardiacPanel Test (e) and a custom hand held assay (f) can inserted to theprototype shown in (c). (g) and (h) Different views of the readerprototype shown in (c).

FIG. 5 shows views of the reader with its two-sided tray design for allfive different ROT types, including, (a) Afla-V aflatoxin ROT (70×27×8mm), (b) BioThreat Alert™ Anthrax ROT (62×30×6 mm), (c) Cardiac PanelROT (80×32×5 mm), (d) a custom hand held assay (70×20×5 mm) and (e)Uni-Gold™ HIV RDT (Trinity Biotech) (39×18×6 mm). Accommodating threeout of five ROT types, the tray lid can be replaced with another one toaccommodate other RDT types without any modifications on the main bodyof the attachment.

FIG. 6 shows a view of the reader functionalities.

FIG. 7A shows one-design-fits-all universal cassette holder.

FIG. 7B shows universal Tray with a spring to accommodate any test thathas dimensions of 85 mm by 35 mm or smaller.

FIG. 7C shows top-view schematics of the tray (left) and side-viewschematics of the tray (right).

FIG. 8 shows reflection and transmission readout modes.

FIG. 9 describes that we have conducted initial measurements to test theperformance of the proposed optical imaging scheme and digital imageprocessing algorithm by analyzing a Bio Threat Alert™ RDT (Tetracoreactivated using highly diluted positive control sample. In order tosimulate the proposed reader platform, we conducted this measurementwith a Samsung Galaxy smartphone mounted on an optical table withadjustable optical and mounting components. For the illumination of RDTunder test, two narrow-band LED arrays (wavelength=565 nm, bandwidth=˜30nm) were used. (a) Prior to acquire RDT image using the proposed opticalscheme, under ambient light, we have recorded a basic smartphone cameraimage in the room conditions, providing no distinct color intensity (onthe test line) that can be identified by human eye. However, in thedigitally processed and enhanced image of the same RDT acquired usingour optical platform (b), digital contrast level has been significantlyimproved between the test line and the background. (c) Although therehas been an analyte-generated background in this RDT image, ourderivative-based filter has corrected this issue, enabling the recoveryof test as well as control line free from the chemical and opticalbackground noise.

FIG. 10 shows exemplary (recorded and automatically processed) images ofnegative (bottom row) and positive (top row) tests for (a) Afla-Vaflatoxin RDT, (b) Cardiac Panel RDT, (c) a custom hand held assaydeveloped by ECBC (Edgewood Chemical Biological Center), (d) BioThreatAlert™ Anthrax ROT, and (e) Uni-Gold™ HIV ROT (Trinity Biotech).

FIG. 11 shows the quantitative calibration curve for the Aflatoxin testthat was generated using 160 different measurements is shown. For eachconcentration level, 2 different RDT were imaged 5 times.

FIG. 12 shows the illumination pattern in the transmission mode

FIG. 13 shows the summary of our measurements on the dry standardOrasure RDT kits which represents various concentration levels rangingfrom blank (0) and negative (N) to high density levels (>5+). In thetable below, the measurement intensity mean, standard deviation and CVvalues are shown together with the corresponding optical density levelsprovided by Orasure.

FIG. 14 shows the schematics for the transmission imaging/readoutmodality using the cell-phone flash and a mirror.

FIG. 15 shows universal reader design that can work with any smartphoneor tablet, utilizing smartphone cases customized for each phone ortablet type.

FIG. 16 shows that, by the optimized selection of the common location offlash/camera for various phones and tablets, the universal readerattachment can work with various smartphones and tablets.

FIG. 17 describes the communication between the smartphone and tablet(application) and the embedded PCB. The reader application running onthe smartphone or tablet can control the flash pulses generated by thecamera and control the electronics on the PCB.

DETAILED DESCRIPTION

A universal rapid diagnostics test reader is disclosed and describedherein that includes a set of control electronics, a digital cameracomponent, an illumination component, a housing component, and a rapiddiagnostics test tray component, wherein the tray can hold at least onerapid diagnostics test having a shape and a size in a fixed positionrelative to the digital camera component and the illumination component,and wherein the reader can accommodate more than one different rapiddiagnostics test.

Many of the deficiencies as outlined earlier are corrected by thecontemplated to embodiments disclosed herein. Specifically, contemplatedembodiments overcome the following limitations:

-   -   Different cassette designs and mechanical dimensions make it        challenging to interface with the imaging system of the reader.        Contemplated embodiments work with a wide range of cassette        sizes, and they avoid the pitfalls of mechanical adapters for        each test type.    -   Conventional readers require sources of illumination and        associated control electronics and battery housed outside the        smartphone. In Mudanyali's conventional reader, the control is        provided by the software application in the smartphone via a        cable which plugs into the smartphone micro USB power connector        or a physical switch that is outside the attachment. External        cabling and physical switches add to the cost and reduce        reliability; besides, many smartphones do not have the        capability for outbound control through their power connector.        Current contemplated embodiments solve this problem with a        wireless control (no wires, cables or physical switches needed).    -   Conventional systems achieve low cost by using a smartphone        which is inserted into a reader body that provides RDT        illumination, ambient isolation, and cassette housing. However,        different models of smartphones from a single manufacturer or        even more from a variety of vendors all have different        mechanical dimensions, and they wouldn't fit into a body        designed for one specific smartphone model, which precludes        users from using their own smartphone for the reader: they have        to buy another dedicated smartphone which is a significant cost        increase. Current embodiments provide a low cost way to        eliminate this problem and enable the use of a wide variety of        mobile devices including smartphones and tablet PCs.

Architecture

A Universal Rapid Diagnostics Test (RDT) Reader has been developed, isdisclosed herein and is shown in FIG. 1A, that can accommodate multiple(at least 5 different test types with dimensions up to 35 mm×85 mm) RDTswithout any need for mechanical modification or external RDT trays. FIG.1A shows a contemplated universal RDT reader attachment 110 in an open115 and closed 120 position. A contemplated communications device, suchas a cell phone, is not shown in this Figure. In this single-pieceadapter design, a printed circuit board (PCB) 150, as shown in FIG. 1Bis utilized that includes multiple illumination light emitting diodes(LEDs), a replaceable and rechargeable battery, a recharging circuitwith its USB port, and a photo-sensor, which is used to wirelesslytrigger/control the illumination LEDs via the cell-phone application.The PCB with the battery is coupled with or affixed into the adapter.The complete reader attachment assembly is wirelessly press-fit onto asmartphone for easy mounting and demounting.

One objective of the development of contemplated embodiments disclosedherein was to introduce a rugged, wireless (controlled using cellphoneflash via the photo-sensor), lensless (no external lens needed),smartphone based universal RDT reader that can continuously operate overextended hours even in field settings. It should be understood that theplatform is cell-phone independent, such that it can be adapted to anycell-phone device with minor or no mechanical modifications. In thisembodiment, we used an inexpensive and rugged Motorola Defy XT 535smartphone. It includes optical and electrical components embedded on aprinted circuit board (PCB) that is powered by a rechargeable battery,which can operate over 12 hours without any need for external power.This contemplated universal reader, without any modification on itsmechanical architecture, accommodates and digitally interprets a broadrange of RDTs to diagnose chemical and biological threats and otherdiseases.

The sensitivity and accuracy of the platform was demonstrated byconducting repeated measurements on positive (including the onesactivated by highly diluted positive control samples) and negativetests. Through a custom-developed smartphone application, the integratedsmartphone-based reader labels digitally processed test results withspatiotemporal information and transfers them to central data collectionpoints (servers) that can be accessed locally and globally.

This smartphone-based RDT reader and spatiotemporal threat/diseasemonitoring platform utilizes a compact snap-on smartphone attachment 210that can be repeatedly attached/detached at the back 295 of thesmartphone devices 290 (see FIGS. 2A and 2B) to acquire digital imagesof RDTs. This scalable attachment is designed to fit differentsmartphone devices (e.g., iPhone or Android phones and tablets) and canbe simply adapted with minimal or no engineering.

The mechanical body of this independent reader attachment is designed tobe robust and easy-to-handle by the user and initially prototyped usinga 3-D printer, which uses ABSplus™ modeling material, a recyclable andeco-friendly thermoplastic. For volume manufacturing using differenttechniques (e.g., injection molding or casting), other material typeswith different material properties can be used. This snap-on readerattachment utilizes inexpensive optical components and printed circuitboards with various electrical components i.e., multiple LEDs (lightemitting diodes) and/or LED arrays, a photo-sensor interface towirelessly control and trigger the illumination LEDs (via the cell-phoneapplication), rechargeable battery as well as a recharging circuitry andits USB port embedded on the same. (see FIGS. 2C and 2D). In FIG. 2C, acontemplated device 210 is shown with a USB connector 220, a “slip-free”grip design 230, a lens assembly 240, protection glass 250 and an “errorproof” multi-track system 260. In FIG. 20, the back of a contemplateddevice 210 is shown with the USB port or connector 220, a rechargingcircuit 221, a battery holder 222, a LED driver circuit 223, an audioport 224 and at least one LED bar 225. Please note that this readerattachment does not require the use of any external lenses for themagnification/demagnification of the RDT image, however it has anoptional tray that allows the user to utilize an additional imaging lens(optional) to meet to requirements of different imaging conditions.

Another important design consideration is the choice of ROT illuminationscheme that has a significant effect on the sensitivity and accuracy ofthe test interpretation. Embedded on a single PC board, multiplediffused LEDs can be used to illuminate (with an illumination angleclose to normal incidence) the RDT under test (see FIG. 2D) in bothreflection and transmission modes (butt-coupled to the ROT under test).The PC board can also a high-power UV led for the digital evaluation offluorescent tests. Unlike reflection and transmission mode LEDs, UV ledis positioned with angle of ˜30-50 degrees to excite the tests ofinterest. Powered by an embedded rechargeable battery, LEDs arewirelessly controlled by a photo-sensor mounted on the PCB. Thesmartphone application, based on the user's selection, digitallyconfigures and utilizes the flash of the cell-phone to trigger thephoto-sensor and control the illumination LEDs automatically duringtesting. This wireless and digital communication with the illuminationLEDs allows the software application to turn on the LED arrays onlyduring image acquisition and observation, decreasing the powerconsumption for extended battery life.

Furthermore, as shown in FIG. 2D, the PCB board also has acustom-designed circuitry with an integrated USB port or connector torecharge the battery of the reader attachment without any need foradditional equipment, simply by sharing the smartphone USB cable withinthe base mechanical body. Moreover, by using this external battery, acontemplated ROT reader is able to evaluate >1000 RDTs and operatelonger than 12 hours without any need for external battery andrecharging. Applying <10 second exposure for image acquisition per test,this enables rapid readout of stacked RDTs without an interruption.

Rather than using broad-band light sources (e.g. ambient light) withvarying intensity profiles and optical spectrum, the use of special LEDillumination can significantly increase the contrast between thecontrol/test lines and the background on RDT images, provided that thewavelength of the illumination is optimized based on spectralmeasurements. For instance, test pads (e.g., nitrocellulose membrane) ofmost commercially-available RDTs produce distinct color signal by theimmobilization of colloidal gold-labeled antigen (e.g., analyte) andantibody (e.g., binding proteins) complexes, exhibiting similar spectralproperties. Based on our initial tests on various RDTs in the market,the use of LEDs with a peak wavelength of between 520 and 590 nmprovides the highest contrast if colloidal gold nano-particles are usedto label target antigen or antibody (in both reflection and transmissionmodes). It should be noted that this optimum center wavelength wasdetermined based on spectral measurements on colloidal gold-based RDTsthat are widely commercially available. Together with thecustom-developed image processing algorithm that is discussed later,optimization of the illumination wavelength has primary importance onthe trans-visual sensitivity of this smartphone based reader platform.

In parallel to the proposed LED illumination scheme, the use of theflash light was examined as an additional illumination source that isalready available in the camera smartphone devices (see FIG. 2F). FIG.2F shows a lens assembly 240 wherein two different systems aredisclosed: a flash system 242 having a diffuser 243 and a lens 244 or aLED system 246 having a lens 244, whereby one of these systems isinstalled in the lens assembly 240. Locating a band-pass filter(optional) that is butt-coupled to the flash light, flash can bealternative to the LED illumination that has been described above (inboth reflection and transmission modes). Although the use of externalLEDs has significant advantages, such as high power output, opticaldesign flexibility and illumination uniformity, implementing the cameraflash light on the RDT reader, as an option for less demandingapplications, can eliminate the need for using LEDs, electricalcomponents, and external battery. In reflection mode, by simply locatinga band-pass filter (e.g., with 520-590 nm pass-band), the flash lightcan replace the reflection mode LEDs. On the other hand, flash light canbe also used for the transmission mode imaging. A reflection elementsuch as a mirror can be placed at the top wall of the attachment toreflect the image of the RDT that is positioned in front of the flashfor transmission mode imaging. Reflected image of the RDT can be thenimaged by the cell phone camera (see FIG. 14).

Cassette Tray

Enclosing the optical imaging interface, the mechanical body of thereader attachment will also ensure the isolation to the RDTs that areloaded to the smartphone reader attachment as shown in FIGS. 1 and 3.Prior to digital evaluation, an RDT is press-fit into a unique cradle onthe tray in an easy, reliable, and error-proof operation. On theinterior side of the universal tray, a series of cradles are formed byridges at different heights, orientations and lateral extensions, asshown in detail in FIG. 3, for 7 different RDTs. A computer-aidedoptimization and design methodology used to design this universal trayis akin to solving a three-dimensional puzzle and can be readily appliedto a different set of RDT shapes and sizes.

Without any modification on the base reader attachment shown in FIG. 2E,multiple trays can be used that are capable of holding at least 5 kindsof RDTs each. FIG. 2E shows a contemplated device 210 where a smartphone290 is connected having a CMOS or complementary metal-oxidesemiconductor 292 and a lens 294. An additional lens 245 is shown, withthe protective glass 250 and the smart track or multi-track system 260.The PCB 275 is also shown. A RDT tray 280 and the tray lid 285 is alsoshown. As a matter of fact, this innovative design eliminates the needof using an individual customized tray for each RDT type andsignificantly decreases the material cost and logistical problems,enabling ease-of-use even in the field settings. By the optimization ofthis innovative approach, the mechanical interface (tray) can bedesigned to accommodate different and larger groups of RDTs. It is alsoimportant to underline that this universal tray fully encloses the RDTsof interest (see FIG. 2A-F) to tackle potential ambient light leakageinto the optical attachment. Since RDT material/packaging may behave asa waveguide that couples the ambient light to the optical imaginginterface, this universal tray design is a vital design feature toensure the repeatability of the measurements.

Moreover, the proposed RDT reader attachment will have a physicalopening (i.e., field-of-view) of ˜45 mm×85 mm to accommodate thisuniversal tray carrying a wide range of RDTs (see FIG. 3A-G). FIGS. 3Athrough 3G show various RDTs 330 that are coupled with the RDT readerattachment 310, wherein the ROT reader attachment is shown in a sideview (upper view) and an above perspective (lower view) in each Figure.Users can rapidly replace an already evaluated RDT with a new one to betested without having any mechanical difficulty, thus enabling testingof large number of stacked RDTs in a short time.

Also, it also allows the user to acquire images of other objects ofinterest, such as user ID card and RDT pouch with type/lot numbers,while the universal RDT tray is retracted from the base attachmentassembly. Digitally linked to the test results, these additional imagescan be processed to extract the relevant identification and securityinformation. Note that the unconventionally wide field-of-viewintroduced here to accommodate a broad range of RDTs provides anopportunity to acquire images of even larger RDTs with larger dimensionsor non-planar packaging (i.e. urine cup) by partially sacrificing thecompactness of the reader attachment. An embodiment of these designprinciples is shown in FIGS. 4-6.

FIG. 4 shows (a, b, c) Photo of the Rapid Test Reader prototypes 410fabricated using a 3-D printer. (a) A prototype with a slip-on universalRDT tray can accommodate up to 7 different RDTs (b) A single-piecedesign that with relatively smaller dimensions can accommodate and fullyenclose up to 7 different RDTs (c) A third single-piece design that canfully enclose and accommodate up to 5 different RDTs has significantlysmaller dimensions and weight as well as smooth edges and corners. (d,e, f) Different types of RDTs, e.g., Uni-Gold™ HIV RDT (d), CardiacPanel Test (e) and a custom hand held assay (f) can inserted to theprototype shown in (c). (g) and (h) Different views of the readerprototype shown in (c).

FIG. 5 shows views of the reader 510 in an open embodiment showing itstwo-sided tray design for all five different RDT types 530, including,(a) Afla-V aflatoxin ROT (70×27×8 mm), (b) BioThreat Alert™ Anthrax RDT(62×30×6 mm), (c) Cardiac Panel RDT (80×32×5 mm), (d) a custom hand heldassay (70×20×5 mm) and (e) Uni-Gold™ HIV RDT (Trinity Biotech) (39×18×6mm). Accommodating three out of five RDT types, the tray lid can bereplaced with another one to accommodate other RDT types without anymodifications on the main body of the attachment.

FIG. 6 shows a view of the reader 610 with a display of the main menu offunctionalities as shown on the smartphone 690 attached.

An alternative contemplated design for a universal cassette holder 710is shown in FIG. 7A. In this contemplated embodiment, the position ofthe cassette 730 is fixed by pushing it against an L-shaped ridge 720;the cassette is kept in place by one or more leaf springs 725. Byleaving the two sides of the cassette unconstrained, this design canaccommodate a wide variety of shapes and sizes with a single design.This kind of flexibility is a huge advantage for the manufacturer andthe users. Rather than just pushing the cassette into the only cradle itwould fit, now the operator must make sure that the cassette fits snuglyin the corner of the L. Also, for a triangular or other non-rectangularshapes the test strip is at an angle but this is easily handled by thesoftware (application running on the cell-phone). Moreover, thisflexible approach, without any hardware and software modifications, mayallow the user to work with emerging next-generation technologies (flowthrough tests) that will be available at the markets in the near future.

In addition to the designs shown in FIG. 1A and FIG. 7A, a completelyuniversal and user-friendly tray design 710 has been developed forcontemplated readers (FIG. 7B). The designs are based on the conceptsshown in FIG. 7C: by constraining the position of the RDT 730 in oneplane only by an L-corner 720 and in the perpendicular direction by aslanted or curved side (727 in FIG. 7C) it is possible to accommodate awide range of RDT shapes and sizes in the same tray. This designutilizes another contemplated flat tray with a spring 725 placed on theside wall to keep the cassettes, which can simply slide into the tray,in place. In this design, the tray has only three walls with an openingat the top for the user to slide in the test cassettes of interest. Theposition of the cassette is fixed by pushing it against to the side ofthe tray; the cassette is kept in place by one or more leaf springs.This design allows the user to work with any cassette type that issmaller than 85 mm by 35 mm. By simply modifying the tray (testinterface, not the main body of the attachment), these dimensions can befurther increased. On the other hand, operator inserts the cassettesinto this mechanical interface by sliding them in all the way into thetray such that ideally the cassette should fit in the L-corner. However,unlike the drawback of the design in FIG. 7A, the potential shift due tothe failure of the operator will be only in longitudinal direction—notin other directions. Our smart cell-phone application will recognizesuch vertical shifts on the position of test casettes and eitherdigitally compensate for it and evaluate the test or warn the operatorto correct the position of the test cassette. Alternatively, a secondspring could be implemented on the fourth side of the tray to ensurepositive contact with the L-corner. To keep the cassette from fallingout of its position, a third spring or springs could be mounted on thetop of RDT. Alternatively, at least one of the tray sidewalls could beslanted 727 or curved as shown in FIG. 7C; note that different cassettethicknesses 730 all fit in with only a slight lateral displacement. InFIG. 7C, a top-view 702 and a corresponding side-view 704 is shown.

Readout

A contemplated reader 810, like the conventional readers disclosedearlier, has three readout modes: fluorescent (not shown), reflectionmode 820 and transmission mode 830, as shown in FIG. 8. The reflectionmode 820 is used in all available readers as it obviously parallelsvisual readout, it is easy to implement, and provides comparableresults. FIG. 8 shows the cell phone camera 891, the cell phone cameralens placement 892, an external imaging lens 893, an orasure testmembrane 894 and a semi-transparent orasure cassette 896, as they areplaced or as they are relative to the reader 810. Conventional readersare used more to avoid operator errors and obtain data in the electronicformat rather than to improve the sensitivity. In the implementation,however trans-visual (better than visual) sensitivity was obtained evenin the reflection mode by virtue of ambient isolation and optimizationof illumination, as described previously. A qualitative example 905 oftrans-visual performance is shown in FIG. 9 where the reader is clearlyable to detect a weak line invisible to the naked eye by using an imageprocessing algorithm similar to conventional readers. An optimized imagefor all five RDTs used in this work is shown in FIG. 10. FIG. 10 showsexemplary (recorded and automatically processed) images of negative(bottom row 1015) and positive (top row 1025) tests for (a) Afla-Vaflatoxin RDT, (b) Cardiac Panel RDT, (c) a custom hand held assaydeveloped by ECBC (Edgewood Chemical Biological Center), (d) BioThreatAlert™ Anthrax RDT, and (e) Uni-Gold™ HIV RDT (Trinity Biotech).

To demonstrate the accuracy of this reader in the reflection mode, astatistically significant number of tests were performed on one highquality RDT cassette capable of quantitative performance, Afla-Vaflatoxin RDT. The result is shown in FIG. 11. In reflection mode, inaddition to the LEDs located on the PCB, cellphone flash can be used toilluminate the RDT under test.

Transmission mode readout was first proposed by Mudanyali et al[18 ] andqualitatively demonstrated to be an alternative to the reflection mode.However, their transmission mode required that both sides of the LFIstrip be open and accessible to light-one side toward the illuminatingsource and the other side toward the camera- and this turned out to be amajor impediment for practical use. The fact is that an overwhelmingmajority of RDT cassettes today on the market has only one open windowfor the strip with the other side being covered by a plastic back (forexamples see FIGS. 4 and 5), thus precluding the use of the transmissionmode except for a small number of specialized cassettes or cassettescustom designed for this mode.

The key insight that led to the contemplated embodiments disclosedherein is that nearly all RDT cassettes on the market are made of whiteor lightly colored plastic that is sufficiently translucent to allowsufficient light transmission through the cassette wall to provideadequate illumination of the LFI strip and ultimately detection by thecamera. In addition, the translucent plastic acts as a diffusersubstantially improving the uniformity of the strip illumination.

The advantages of these contemplated embodiments are summarized asfollows:

-   -   The signal captured by the camera contains information about the        density of gold particles throughout the thickness        (3-dimensional morphology) of the paper strip rather than just        on the surface or close to the surface, which will substantially        increase the lower detectable limit compared with the reflection        mode measurement.    -   Translucent plastic in the cassette wall in the pathway of light        is generally strongly scattering and diffuses light which        contributes to the uniformity of the illumination of the strip        and this minimizes measurement errors.    -   The reflection mode illumination may cause multiple reflections        from the RDT cassette, causing trouble in digital processing        steps. This is particularly the case when the cassette window is        covered with plastic, which is the prevailing situation with        rapid tests used with saliva and which often also have weaker        signal. The transmission mode avoids these problems.    -   Because of the above all light reaching the camera is diffused        either through two layers of plastic with and without the strip        or one layer plus the strip. Consequently the contrast over the        camera field of view is considerably more uniform than for the        reflection case as shown below, which reduces problems with        camera saturation and white balance variation.    -   For the same reason as above, there are no shadows around the        walls of the window (due to the oblique illumination angle of        the light sources in reflection mode) which are also detrimental        for the measurement. In fact, it was observed that the backside        illumination creates a sharp edge 1245 around the window which        may be beneficial in determination of the window position and        line edges (FIG. 12)    -   Blank (not-activated) tests (RDTs or LFIs) have test and control        bands. Antibodies or other chemicals that are necessary for the        color changes are dispensed (e.g., injected or coated) to these        bands and required for the successful operation. In transmission        mode, these bands can be screened before the use as a quality        control mechanism at the manufacturing side.

The performance of this transmission mode was confirmed using a set ofcalibration test RDTs from Orasure. The results are shown in FIG. 13.They confirm trans-visual sensitivity of <0.5% Optical Density andexcellent CV of ˜1%.

The transmission mode LEDs are located at the tray (behind the RDT undertesting), and butt-coupled to the back of the plastic cassette.Moreover, instead of external LEDs, cell-phone flash can be usedprovided that a mirror is located at the tray. In thisflash-transmission geometry, the RDT is located on the top of the flashand parallel to the cell-phone camera. The flash is controlled by thecell-phone application and illuminates the back of the cassette. Themirror located at the top of the tray reflects the image which isrecorded via the cell-phone camera. See FIG. 14 for the transmissionmode readout geometry using flash. In FIG. 14, the top view 1407 of acontemplated reader 1410 is shown in transmission readout geometry 1430,wherein the cell phone 1490 and the cell phone camera module 1493 isshown with an optional external lens 1496. The attachment point 1415 forthe reader 1410 is shown, along with a mirror 1444 and the RDT 1431 nearthe cell phone flash 1491.

It should be emphasized that the all five imaging/readout capabilitiesdescribed here, namely, (i) Reflection mode readout using the LEDsembedded on PCB, (ii) Reflection mode readout using cell-phone flash,(iii) Transmission mode readout using the LEDs embedded on the RDT tray(door), (iv) Transmission mode readout using the cell-phone flash withthe use of a mirror and (v) Fluorescent mode readout using one or moreLEDs on the PCB can be implemented on the same platform with minor or nomechanical adaptation or changes. The PCB has been designed to operateat any of the readout modes. Wirelessly controlling the PCB by sendingflash pulses, the smartphone application allows the user to switchbetween readout modes or automatically chooses the readout mode based onthe RDT type.

Smartphone Options

All of the currently-used, conventional reader implementations requiremultiple mechanical attachments that are physically customized to fitonto different smartphones, which may be acceptable for higher endprofessional markets, where users want to buy a complete readerinstrument and are willing to pay for the cost of the smartphone inaddition to the cost of the attachment, but it does limit the size ofthe addressable market and it is definitely too expensive for consumermarkets. Different phones can be fitted on the same reader body byhaving adjustable rails or hooks but these tend to be clumsy, expensive,and they can be misadjusted through use.

Contemplated embodiments disclosed herein implement a low costadaptation layer by using smartphone cases. These protective cases arevery popular and sold in large quantities. Because of the huge volume,simple design, and little material they are very inexpensive ($10 to$30) and by definition they fit the smartphone 1590 perfectly. Thearchitecture of this solution is shown FIG. 15. Case 2 1536 is affixedto the reader body 1510 via the attachment 3 1537. The cassette tray 51521 is also shown. This can be screws, glue, double sided tape, Velcrotape or another thin mechanical attachment or adaptor; each with itsadvantages or disadvantages but all feasible. The attachment would bepermanent or semi permanent. It would initially be done in the factoryand later maybe by the user.

With this arrangement reader body 4 can be the same for all smartphonesand rapid tests and can be made inexpensively in large quantities withhard mold injection process but, in order to be universal, it has to besomewhat larger than any contemplated smartphone. The body 1632 mustalso have an opening on the top 1634 to accommodate differentsmartphones with different positioning of the camera 1630 relative tothe main smartphone body 1690 and the flash 1637 relative to the camera1630, which is shown in FIG. 16 in an exaggerated way to illustrate thepoint.

An alternative to the use of commercially available smartphone cases isto design a universal smartphone cradle by following the same principlesas shown in FIGS. 7B and 7C, i.e. fixing the position of the smartphonein a cradle by the use of L-corner, springs, and slanted sides.

Auto-Focus Approach

The unique optical interface of the reader attachment was designed touniformly illuminate the field-of-view of an area of larger than ˜60mm×˜90 mm. This ensures that any rapid test cassette to be analyzed bythis reader will be uniformly illuminated such that the readingvariation caused by the illumination intensity is minimized, increasingthe repeatability of measurements. On the other, the digital focusing ofcell-phone camera is challenging during the image acquisition due to theneed for most uniform illumination on the RDT plane that is located only˜20-60 mm from the cell-phone camera. Live RDT image consists of onlyspatial low-frequency components at this illumination configuration,often causing the camera's auto-focus algorithm to fail. For successfulfocusing by the camera, there should be significant amount of spatialhigh-frequency components (e.g., sharp edges and transitions or lightoscillations) on the live image.

To help the camera achieve better optical focus, we first turn on thecamera's flash in burst mode to create the amount of contrast necessaryto achieve focusing on the image. This non-uniform short point sourceillumination provided by the flash generates the amount of contrastnecessary for such focus algorithms.

Once the camera has focused on the image, we are able to preserve thatfocus distance throughout the test cycle by creating a class whichimplements the Android autofocus Callback interface and setting aBoolean flag upon successful focus. We are able to obtain the pointsource illumination from the camera's flash in order to improve ourfocus while still maintaining an even, single wavelength illuminationfor the image capture. Though our solution was designed to work with acontrast detection auto-focus system, it will improve focus distancedetection for systems which use phase detection algorithms as well.

Illumination Control

Readers require sources of illumination and associated controlelectronics and battery housed outside the smartphone. In Mudanyali'sreader [18], the control is provided by the software application in thesmartphone via a cable which plugs into the smartphone micro USB powerconnector. However, many smartphones do not have the capability foroutbound control through their power connector. On the other hand allsmartphones have an audio jack which can be used to transmit control viaan audio signal. Both approaches work but they do require externalcabling and connectors that add to the cost and reduce reliability.

Contemplated embodiments provide a wireless control connection via RFsignals, including Bluetooth, WiFi, and Near Field Communications (NFC),or using an optical signal generated by the flash in the smartphone1790. The block diagram of this approach is shown in FIG. 17. Uponusers' command to initiate the test 1715, the software (not shown)application generates an Android command that switches the flash 1767on. The burst of the flash light (not shown) is detected by a photosensor (diode or transistor) 1744 that is located on the PCB 1735,wherein the photo sensor 1744 triggers the illumination controlelectronics 1742 to activate the LEDs 1740 for RDT 1730 illumination. Asingle burst is sufficient for the usual operation where theillumination parameters are fixed and only the timing needs to becontrolled; if more complex control is desired (i.e. choice of LEDs,light level, or illumination duration), the flash 1767 can be commandedto generate a sequence of bursts that can be appropriately decoded bythe illumination control 1742. Note that this optical control method notonly avoids any external cables but it is also considerably lessexpensive than other wireless methods that might be considered. Insummary, in contemplated platforms, the flash can be used for reflectionand transmission mode illumination of RDTs as well as the wirelesscontrol of the reader attachment by the cell-phone application.

REFERENCES

The following references are referred to herein by their referencenumber. These references are incorporated herein in their entirety byreference.

1 P. Yager, T. Edwards, E. Fu, K. Helton, K. Nelson, M. R. Tam and B. H.Weigl, Nature, 2006, 442, 412-418.

2 S. Banoo, D. Bell, P. Bossuyt, A. Herring, D. Mabey, et al., Nat RevMicrobio, 2006, 4, 21-31.

3 M. Dhorda, P. Piola, D. Nyehangane, B. Tumwebaze, A. Nalusaji, C.Nabasumba, E. Turyakira, R. McGready, E. Ashley, P. J. Guerin, G.Snounou, Am J Trop Med Hyg, 2012, 86(1), 93-95.

4 I. N. Okeke, R. W. Peeling, H. Goossens, R. Auckenthaler, S. S.Olmsted, J. F. de Lavison, B. L. Zimmer, M. D. Perkins, and K.Nordqvist, Drug Resist Updat, 2011, 14(2), 95-106.

5 C. Drakeley and H. Reyburn, Trans R Soc Trop Med Hyg, 2009, 103(4),333-337.

6 C. K. Murray, R. A. Gasser Jr, A. J. Magill, and R. S. Miller, ClinMicrobiol Rev, 2008, 21(1), 97-110.

7 J. Skarbinski, P. O. Ouma, L. M. Causer, S. K. Kariuki, J. W.Barnwell, J. A. Alaii, A. M. de Oliveira, D. Zurovac, B. A. Larson, R.W. Snow, A. K. Rowe, K. F. Laserson, W. S. Akhwale, L. Slutsker, and M.J. Hamel, Am J Trop Med Hyg, 2009, 80(6), 919-926.

8 L. A Mills, J. Kagaayi, J. P. Shoff, K. Newell, J. B. Bwanika, V.Ssempijja, S. Aluma, T. C. Quinn, S. J. Reynolds, R. H. Gray, Trans RSoc Trop Med Hyg, 2010, 104(3), 237-239.

9http://www.itu.int/ITU-D/ict/publications/idi/material/2012/MIS2012_highlights_short.pdf,Retrieved on 12.25.2012.

10 H. Zhu, O. Yaglidere, T. Su, D. Tseng, and A. Ozcan, Lab Chip, 2011,11, 315-322.

11 D. Tseng, O. Mudanyali, C. Oztoprak, S. O. lsikman, I. Sencan, O.Yaglidere, and A. Ozcan, Lab Chip, 2010, 10, 1787-1792.

12 O. Mudanyali, D. Tseng, C. Oh, S. O. lsikman, I. Sencan, W. Bishara,C. Oztoprak, S. Seo, B. Khademhosseini, and A. Ozcan, Lab Chip, 2010,10, 1417-1428.

13 G. McKiernan, Searcher, 2010, 18 (3), 48-51.

14 A. W. Martinez, S. T. Philips, E. Carrilho, S. W. Thomas III, HayatSindi and G. M. Whitesides, Anal Chem, 2008, 80, 3699-3707.

15 Adler R., Health Care Unplugged: The Evolving Role of WirelessTechnology, California HealthCare Foundation, 2007

16. A. Coskun, J. Wong, D. Khodadadi, R. Nagi, A. Tey, and A. Ozcan, LabChip, 2012, DOI: 10.1039/C2LC41152K.

17 http://www.idc.com/qetdoc.jsp?containerId=prUS23771812.

18 O. Mudanyali, S. Dimitrov, U. Sikora, S. Padmanabhan, I. Navruz andA. Ozcan, Lab Chip, 2012, 12, 2678-2686.

19http://holomic.com/content/2012/07/01/holomic-introduces-a-smartphone-based-rapid-test-reader-at-aacc-2012/Introductionof rapid reader

20 PCT/US2012/040282

21 Faulstich et al in “lateral Flow Imunoassay, Edited by Raphael Wongand Harley Tse, Springer 2009.

22http://www.qiagen.com/about-us/contact/oem-services/ese-instruments/esequant-lateral-flow-system,Retrieved on 12.31.2013.

23 US2013/0244339

Thus, specific embodiments and methods of a universal smartphone-basedrapid diagnostic test reader with trans-visual sensitivity have beendisclosed. It should be apparent, however, to those skilled in the artthat many more modifications besides those already described arepossible without departing from the inventive concepts herein. Theinventive subject matter, therefore, is not to be restricted except inthe spirit of the disclosure herein. Moreover, in interpreting thespecification and claims, all terms should be interpreted in thebroadest possible manner consistent with the context. In particular, theterms “comprises” and “comprising” should be interpreted as referring toelements, components, or steps in a non-exclusive manner, indicatingthat the referenced elements, components, or steps may be present, orutilized, or combined with other elements, components, or steps that arenot expressly referenced.

We claim:
 1. A universal rapid diagnostics test reader, comprising: aset of control electronics, wherein the set of control electronicscomprise processor electronics that control an illumination component; asmartphone, wherein the smartphone comprises a wireless communicationcomponent; a digital camera component, wherein the digital cameracomponent is operatively located in the smartphone, wherein the wirelesscommunication component comprises a wireless connection that isoperatively initiated and engaged by a flash in the digital cameracomponent and a photo-detector in the set of control electronics, andwherein the flash controls an illumination component; an illuminationcomponent, wherein the illumination component comprises at least onelight-emitting diode and a transmission mode of operation where therapid diagnostic test is located between the light-emitting diode andthe camera component; a housing component, and a single rapiddiagnostics universal test tray comprising a series of cradles formed byridges at different heights, orientations, and lateral extensions,wherein the single tray can hold at least two different types or kindsof rapid diagnostics test having a shape and a size in a fixed positionrelative to the digital camera component and the illumination component,wherein the single tray can only hold one test at one time, and whereinthe single rapid diagnostics universal test tray can accommodatedifferent types or kinds of rapid diagnostics tests in the sameuniversal test tray without additional mechanical adaptation to eachtest before it is placed in the tray, without a mechanical adaptersurrounding each test before it is placed in the rapid diagnosticsuniversal test tray, or without additional mechanical components appliedto each test before it is placed in the rapid diagnostics universal testtray.
 2. The universal rapid diagnostics test reader of claim 1, whereinthe digital camera component comprises at least one lens, at least oneimage sensor, at least one analog to digital converter, at least onedigital image processor, at least one flash, at least one microprocessoror a combination thereof.
 3. The universal rapid diagnostics test readerof claim 1, wherein the digital camera component uses autofocus andwherein the illumination component comprises flash illumination.
 4. Theuniversal rapid diagnostics test reader of claim 1, wherein the wirelesscommunication component comprises a wireless connection that isoperatively initiated and engaged by non-optical functions.
 5. Theuniversal rapid diagnostics test reader of claim 4, wherein non-opticalfunctions comprise WIFI, IEEE standard Bluetooth, or Near FieldCommunications.
 6. The universal rapid diagnostics test reader of claim1, wherein the flash controls light-emitting diode selection.
 7. Theuniversal rapid diagnostics test reader of claim 1, wherein thesmartphone is enclosed in its case before being operatively engaged withthe housing component.
 8. The universal rapid diagnostics test reader ofclaim 1, wherein the smartphone is operatively engaged with the housingcomponent.
 9. The universal rapid diagnostics test reader of claim 8,wherein the smartphone is further constrained with an L-shaped cornercomponent.
 10. The universal rapid diagnostics test reader of claim 8,wherein the smartphone is held in a fixed position in a planar directionwith at least one spring.
 11. The universal rapid diagnostics testreader of claim 8, wherein the smartphone is constrained in aperpendicular direction with a slanted side or a curved side.
 12. Theuniversal rapid diagnostics test reader of claim 1, wherein theillumination component includes illumination of rapid diagnostics testby one or more light emitting diodes at the wavelength of imaging forchromatographic rapid diagnostic tests or at the excitation wavelengthfor the fluorescent rapid diagnostic tests.
 13. The universal rapiddiagnostics test reader of claim 12, wherein the illumination componentcomprises a reflection mode of operation where one or morelight-emitting diodes and the camera component that are on a front sideof the rapid diagnostic test with the light-emitting diode axis that isroughly perpendicular to the rapid diagnostic test plane.
 14. Theuniversal rapid diagnostics test reader of claim 13, wherein thelight-emitting diode is the flash light-emitting diode that is part ofthe camera component illuminating a strip on the rapid diagnostics testfrom the front side.
 15. The universal rapid diagnostics test reader ofclaim 1, wherein the rapid diagnostic test may comprise a bare strip, astrip on a translucent plastic backing, a conventional strip packaged ina translucent plastic cassette or a combination thereof.
 16. Theuniversal rapid diagnostics test reader of claim 1, wherein thelight-emitting diode is the flash light-emitting diode that is a part ofthe camera component.
 17. The universal rapid diagnostics test reader ofclaim 16, wherein the camera is imaging a strip via a mirror placed infront of the rapid diagnostics test.
 18. The universal rapid diagnosticstest reader of claim 1, wherein the housing component is designed toenclose all components of the reader into a light tight enclosure andwherein the rapid diagnostic test is illuminated only by theillumination from the reader and not illuminated by ambient light. 19.The universal rapid diagnostics test reader of claim 1, wherein therapid diagnostics tray secures a rapid diagnostics test in the tray withan L-shaped corner component.
 20. The universal rapid diagnostics testreader of claim 1, wherein the rapid diagnostic test is held in a fixedposition in a planar direction with at least one spring.
 21. Theuniversal rapid diagnostics test reader of claim 1, wherein the rapiddiagnostic test is constrained in a perpendicular direction with aslanted side or a curved side.
 22. The universal rapid diagnostics testreader of claim 1, wherein the more than one rapid diagnostic tests areconstrained in their position by their own cradles and posts customizedon the same tray.